Isolation of geniposide in Gardenia jasminoides was accomplished by high performance GF254 TLC and the content of the said component in the herb and its preparation determined by linear scanning at ?248nm. The method was simple and accurate.

Objective To observe the technical feasibility and safety of delta -shaped (DS)anastomosis in totally laparoscopic distal gastrectomy (TLDG).Methods A retrospective analysis of 9 cases underwent TLDG with D2 lymphadenectomy and DS anastomosis.Results All the patients underwent TLDG with D2 lymphadenectomy and DS anastomosis successfully by the same doctor and his team.The total operative time was (178 ±43)minutes,and the DS anastomosis time was (32 ±8)minutes.All the patients achieved microscopic cancer -free margin and the number of lymph nodes harvested was (27 ±8)per patient.The average time to flatus,time to fluid diet were (3.8 ± 1.7)days and (3.9 ±1.8)days respectivly.One patient developed to fat liquefaction of incision,and one developed to urinary tract infection after the operation.No anastomotic bleeding,anastomotic leakage,anastomotic stenosis or other anastomotic complications happened among all patients.Conclusion DS anastomosis is safe and convenient.It is an ideal choice for reconstruction in TLDG with D2 lymphadenectomy.

BACKGROUND: Allicin has been proved to influence the proliferation and apoptosis of LM-8 cells.OBJECTIVE: To study the effects of allicin on the expression of apoptosis protein Bcl-2 and Bax in C3H mouse osteosarcoma cell line LM-8, and the correlation of the morphological changes of LM-8 cells with the changes of Bcl-2 and Bax.DESIGN: Randomized grouping, non-blind, cell level, in vitro study.SETTING: The experiment was carded out from September 2006 to May 2007 at the Center Laboratory of Union Hospital,Tongji Medical College, Huazhong University of Science & Technology.MATERIALS: Osteosarcoma cell line LM-8 was purchased from the Animal Center of the Fourth Military Medical University of Chinese PLA. Allicin presented by Wuhan Huihai Co., was a sulphide isolated from garlic corn. Cell Counting Kit-8 (CCK-8) from Dojindo Laboratories; Bcl-2 antibody, Bax antibody and SP immunohistochemistry kit from Fuzhou Maixin Biotech Co.METHODS: LM-8 cells were cultured in RPMI 1640 culture medium, then cell climbing films were made, and SP immunocyte-histochemistry was used to detect the expression of Bcl-2 and Bax protein. Inverted microscope was performed to observe the morphologic changes of LM-8 cells before and after adding with 5.0, 10.0 and 15.0 mg/L allicin. Serial subcultivated LM-8 cells were adjusted to 7.5× 107 L-1 in concentration, added to 96-pore culture plate, and treated with allicin at a serial concentration of 1.0, 5.0, 10.0 and 15.0 mg/L. Additionally blank culture medium pores without cells and allicin and pores without allicin were taken as control groups. The 96-pore culture plate was maintained for 24, 48 and 72 hours, and then fetched. CCK-8 was added to determine the optical density value. The inhibition rate of LM-8 cell growth was also calculated. LM-8 cells were treated with different concentrations of allicin (5.0, 10.0 and 15.0 rag/L) for 24, 48 and72 hours, and then we carried out an apoptosis analysis by flow cytometry.MAIN OUTCOME MEASURES: Expression of Bcl-2 and Bax protein in LM-8 cells; cell morphous, cell proliferation,and cell apoptosis.RESULTS: ①Expression of Bcl-2 and Bax protein: Allicin could lower the expression of Bcl-2 protein and enhance the expression of Bax. The positive expression of Bcl-2, Bax and Bcl-2/Bax expression all showed significant differences compared with control group (P ＜ 0.01).②LM-8 cell morphous: LM-8 Cells treated with allicin presented typical apoptosis changes under microscope.③LM-8 cell proliferation: Allicin could inhibit the growth of osteosarcoma cell line LM-8, when concentration of allicin added from 1.0 mg/L to 15.0 mg/L, the inhibition rate of LM-8 cells at 72 hours increased from (23.87±3.26)% to (58.32±5.38)%, and 50% inhibiting concentration was 11.09 mg/L.④LM-8 cell apoptosis: After 72 hours with treatment of allicin of 5.0, 10.0 and 15.0 mg/L in concentration, apoptosis rate of LM-8 cells revealed a dose-dependent relationship (P ＜ 0.05).CONCLUSION: ①Allicin can inhibit the proliferation of LM-8 cells in a dose-time-dependent manner.②Allicin can induce the apoptosis of LM-8 cells in a dose-dependent manner.③Allicin can inhibit LM-8 cell proliferation and induce LM-8 cell apoptosis, which is related to down-regulate Bcl-2 protein expression and up-regulate Bax protein expression.

BACKGROUND: The implantation technique of bone marrow mesenchymal stem cells (MSCs) is of important significance for repair of brain injury. However, its action pathway still needs to be investigated.OBJECTIVE: To observe the changes of Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus of rats with cerebral ischemia in which MSCs were implanted, and to analyze the action mechanism of intracranial implantation of MSCs inhibiting neuronal apoptosis.DESIGN: A randomized controlled experiment.SETTING: Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College.MATERIALS: Twenty-four male adult Wistar rats, weighing 180 to 240 g, were provided by the Experimental Animal Center of Shandong University. The involved rats were randomized into 3 groups with 8 in each: control group, injury group and implantation group. 5-bromodeoxyuridine (BrdU, Sigma Company), TUNEL kit, Bcl-2, Bax antibody kit were purchased from Beijing Zhongshan Bioengineering Company.METHODS: This study was carried out in the Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College between April 2005 and September 2006. Rats in the injury group and implantation group were developed into rat models of cerebral ischemia by suture of external carotid artery. Seven days later, the successful rat models in the implantation group were injected in the cerebral cortex and striatum with 2×1012 L-1 MSCs suspension primarily cultured in vitro. The processing of the experimental animals corresponded to the requests of Animal Ethics.MAIN OUTCOME MEASURES: Neuronal apoptosis and Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus were detected by TUNEL staining and immunohistochemical method on the 7 and 14 days after successful modeling, separately.RESULTS: ①Neuronal apoptosis: On the 7th day after successful modeling, apoptotic cells were not found in the control group, and apoptotic cells in the implantation group were significantly fewer than those in the injury group (P ＜0.01). ② Bcl-2 and Bax expressions: On the 14th day after successful modeling, Bcl-2-positive neuronal expression in the cerebral cortex and hippocampus of rats in the injury group was significantly weaker than that in the control group and implantation group (P ＜ 0.01). Bax-positive expression in the cerebral cortex and hippocampus of rats in the injury group was significantly stronger than that in the control group and implantation group (P ＜ 0.01).CONCLUSION: MSCs can promote Bcl-2 expression and inhibit Bax expression of rats with cerebral ischemia injury,and accordingly neuronal apoptosis will be reduced.

0. 05). This paper surggests that the activity of Na+-K+ ATPase on erythrocyte membrane may be a indicator in the diagnosis of stroke and human senility. The mechanism of changes of Na+-K+ATPase was also discussed in this paper.

Objective To investigate the pathogen of 21 infective endocarditis (IE) cases treated with operation in Shanghai Children's Medical Center from 2007 to 2010.Methods Blood culture,vegetation culture and vegetation PCR assay(target gene to the conserved region V3 in 16SrRNA gene) were detected in 21 IE patients; multiplex PCR amplification of staphylococci for methicillin-resistant staphylococcus was performed.Results Of 21 IE cases,20 cases were detected positive by vegetation PCR with the detection rate of 95.2％,12 IE cases were detected positive by blood culture with the detection rate of 57.1％,2 IE cases were detected positive by vegetation culture with the detection rate of 9.5％.The difference of the positive rates of the three methods was statistically significant (P ＜ 0.0001).The vegetation PCR of one case was actinobacillus actinomycetemcomitans,while the blood culture was haemolysis pasteurell which was inconsistent with the vegetation PCR result.Howerver,the PCR result of colony obtained by blood culture was consistent with vegetation PCR that was confirmed as actinobacillus actinomycetemcomitans.The endocardium PCR results of 11 IE cases were consistent with the results of blood culture.MecA gene was detected by multiplex PCR,which could identify methicillin-resistant staphylococcus quickly,sensitively and accurately and could also effectively identify methicillin resistant staphylococcus aureus,when coupled with femA gene detection,thus glycopeptides antibiotic could be prescribed promptly.All the 21 patients recovered and discharged without infection recurrence in the follow-up.Conclusion Universal primer V3 coupled with multiplex PCR can improve vegetation pathogen detection rate of IE patients and is minimally influenced by antibiotic therapy.Multiplex PCR can be applied for etiological diagnosis of IE patients with indication of surgery and negative blood culture or difficult diagnosis,contributing to post-surgery antibiotics selection and improvement of recovery rate of IE patients.

Objective To investigate the predisposing alleles of HLA-DR genes in pemphigus. Methods Polymerase chain reaction specific sequence primers (PCR-SSP) method was applied to type HLA-DR subregion in the patients with pemphigus vulgaris (PV), pemphigus erythematosus (PE) and matched control subjects of Han nationality from Jiangsu and Anhui provinces. Results The results demonstrated that DR4, DRB1*14 (*1401, *1404,*1405)gene frequencies were significantly higher in both PV(Pc

BACKGROUND: It has reported that nicotinamide is capable of protecting intervertebral disc (IVD) against interleukin-1β (IL-1β) or tumor necrosis factor-alpha (TNF-α) induced degeneration. However, the protective mechanism of nicotinamide on IVD cells apoptosis and proliferation remains unclear.OBJECTIVE: To investigate regulatory effects of nicotinamide on rabbit nucleus pulposus cell apoptosis and proliferation in vitro.DESIGN, TIME AND SETTING: Randomized control grouping design, which was carried out in the Laboratory of Orthopaedics and Stem Cell Center, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology from April to October 2007.MATERIALS: Ten Japanese white rabbits (aged 2-3 months weighing 1.5-2.0 kg) were used in this study. Furthermore, nucleus pulposus cells obtained from L1-6 lumbar spine were harvested and cultured for further experiments.METHODS: The NP cells were divided into 6 groups, including control group (without any drug as control), nicotinamide group (0.5 g/L nicotinamide), IL-1β group (10 μg/L IL-1β), IL-1β + caspase group (10 μg/L IL-1β and non-specific caspase inhibitor Z-VAD-FMK), IL-1β + small-dose nicotinamide group (10 μg/L IL-1β and 0.05 g/L nicotinamide), and IL-1β + large-dose nicotinamide group (10 μg/L IL-1β and 0.5 g/L nicotinamide). After 3 days of culture, the cells were examined with Annexin V-PI staining, caspase-3, 8 and 9 activity staining and MTT assay.MAIN OUTCOME MEASURES: The apoptotic rates, the positive rates of caspase-3, 8 and 9 activity staining and the absorbance of MTT assay of each group.RESULTS: ① As compared to IL-1β group, the apoptotic rates were decreased in the IL-1β + caspase group and IL-1β + large-dose nicotinamide group (P < 0.01). ②As compared to IL-1β group, the positive rates of caspase-3, 8 and 9 activity staining were decreased in the IL-1β + caspase group, IL-1β + large-dose and small-dose nicotinamide groups (P < 0.01 or P < 0.01). ③As compared to IL-1β group, the absorbance was increased in the IL-1β + caspase group and IL-1β + large-dose nicotinamide group (P < 0.01).CONCLUSION: Nicotinamide is capable of promoting cell proliferation and inhibiting IL-1β induced apoptosis of nucleus pulposus cells in vitro. The inhibition of apoptosis mainly acts via inhibition of the mitochondrial pathway.

Objective To investigate the relationship between glycated albumin ( GA ) and extent of coronary lesions, GRACE score in patients with acute non?ST segmentelevation myocardial infarction ( NSTEMI) . Methods A total of 226 NSTEMI patients who successfully underwent coronary angiography ( CAG) were enrolled in the study. Groups:( 1) According to GA level,the patients were divided into 3 groups:GA<14. 2% group,14. 2%≤GA≤17. 0% group,GA>17. 0% group. ( 2) According to the extent of coronary le?sions,the patients were divided into 2 groups:single or double branch lesion group,three and/or left main lesion group. ( 3) According to the GRACE score,the patients were divided into 3 groups:Low?risk GRACE score≤108 points group,Medium?risk 108 points140 points group. The extent of coronary lesions was evaluated by Gensini score. The clinical characteristics and Gensini score,GRACE score of each group were compared. Pearson/Spearman correlation analysis and logistic regression were used to analyze the association of GA with the severity of coronary artery disease and GRACE score. Results With glycated albumin increasing,the Gensini score(56. 51±38. 57,68. 30±35. 57,77. 38±36. 52),GRACE score(139. 43±29. 96,149. 77±38. 33,170. 75±27. 52) increased significantly,and significant differences were found between groups( F=5. 587,16. 006,P=0. 004,0. 000) . The ejection fraction( EF) of 3 groups were signif?icantly decrease((58. 30±13. 95)%,(56. 45±10. 79)%,(53. 06±12. 51)%;F=3. 126,P=0. 046). Proportion of severe coronary lesions of 3 groups were increase significantly( 59. 5%( 44/74) ,68. 2%( 60/88) ,87. 5%( 56/64),χ2=13. 528,P=0. 001). The level of GA in three and/or left main lesion group was higher than that in sin?gle or double branch lesion group((13. 92±3. 14)% vs. (16. 80±3. 58)%,t=-5. 693,P=0. 000). The level of GA in High?risk group was higher than that in Low?risk group ( ( 14. 70 ± 1. 54 )% vs. ( 16. 63 ± 4. 02 )%, t=6. 512,P=0. 002) . Correlation analysis showed that the level of GA had significant positive correlation with Gensini score and GRACE score(r=0. 309,0. 265;P=0. 000,0. 000),while had a negative correlation with LVEF(r=-0. 149,P=0. 034). Logistic regression analysis indicated that GA was independent risk factors for severity of coronary artery disease in patients with NSTEMI who successfully underwent CAG( OR=1. 441,95%CI:1. 160?1. 790,P=0. 001) . Conclusion GA level is increase in NSTEMI patients with severe coronary ar?tery disease and risk stratification high. GA is the independent risk factors for severity of coronary artery disease in patients with NSTEMI;GA has significant correlation with dangerous degree in patients with NSTEMI.

Objective To study the clinical characteristics, treatment and prognosis of infective endocarditis in children. Methords Clinical data from 83 patients of infective endocarditis admitted from 1998 to 2012 were retrospectively analyzed. Results In a total of 83 patients, there were 53 males and 30 females, and the average age was 6.8±4.6 years. The main clinical characteristics were fever (77.1%) and mild to moderate anemia (71.1%). The C-reaction protein (67.5%), erythrocyte sedimen-tation rate (60.2%), and white blood cell (47.0%) were elevated. Twenty (24.1%) patients had embolism. Blood culture was pos-itive in 56 (67.5%) cases with bacteria mainly being Gram-positive and Streptococcus and Staphylococcus accounted for 89.3%. Vancomycin and other sensitive antibiotics were effective. Neoplasm was detected in 68 cases (82%) by transthoracic echocar-diograerphy. Fifty-ifve (66.2%) patients underwent cardio surgery. Seven patients (8.4%) died. Conclusion In recent years, the distribution of pathogenic bacteria in infective endocarditis had changed. Streptococcus mitis and Staphylococcus aureus has become a major pathogens and need to be treated by vancomycin and other sensitive antibiotics. The detection rate of neoplasm is higher by echocardiography.